Granted Patent B2

US 12,220,455 · App. 18/345,893 · Granted Feb 11, 2025

Inventors: Sean Michael Sullivan (Escondido, CA); Daiki Matsuda (San Diego, CA); Kiyoshi Tachikawa (San Diego, CA); Padmanabh Chivukula (San Diego, CA); Priya Prakash Karmali (San Diego, CA); Jared Henry Davis (Poway, CA); Yanjie Bao (San Diego, CA); Amit Sagi (San Diego, CA)

Assignee: Arcturus Therapeutics, Inc.

A61K39/215A61K9/5123A61K39/12A61K47/10A61K47/20A61K47/26C07K14/005C07K14/1808C12N7/00C12N15/86A61K38/00A61K2039/53C12N2770/20022C12N2770/20034C12N2770/36122C12N2770/36134C12N2830/42C12N2830/50

Abstract

Claims (59)

1. A nucleic acid molecule comprising:

(i) a first polynucleotide encoding one or more viral replication proteins, wherein the first polynucleotide is codon-optimized as compared to a wild-type polynucleotide encoding the one or more viral replication proteins, and wherein the first polynucleotide comprises a sequence having at least 80% identity to the sequence of SEQ ID NO:72; and

(ii) a second polynucleotide comprising a transgene encoding an antigenic protein or a fragment thereof, wherein the antigenic protein is a coronavirus protein encoded by a sequence having at least 90% identity to the sequence of SEQ ID NO: 122.

2. The nucleic acid molecule of claim 1 , further comprising a 5′ untranslated region (UTR), wherein the 5′ UTR comprises an alphavirus 5′ UTR sequence.

3. The nucleic acid molecule of claim 2 , wherein the 5′ UTR comprises a sequence of SEQ ID NO:73, SEQ ID NO:74, or SEQ ID NO:75.

4. The nucleic acid molecule of claim 1 , further comprising a 3′ untranslated region (UTR), wherein the 3′ UTR comprises an alphavirus 3′ UTR sequence.

5. The nucleic acid molecule of claim 4 , wherein the 3′ UTR comprises a poly-A sequence.

6. The nucleic acid molecule of claim 1 , wherein the first polynucleotide is located 5′ of the second polynucleotide.

7. The nucleic acid molecule of claim 6 , further comprising an intergenic region located between the first polynucleotide and the second polynucleotide.

8. The nucleic acid molecule of claim 7 , wherein the intergenic region comprises a sequence having at least 85% identity to a sequence of SEQ ID NO:77.

9. The nucleic acid molecule of claim 1 , wherein the nucleic acid molecule is

(a) a DNA molecule; or

(b) an RNA molecule, wherein T is substituted with U.

10. The nucleic acid molecule of claim 9 , wherein the DNA molecule further comprises a promoter located 5′ of the 5′ UTR, wherein the promoter is a T7 promoter, a T3 promoter, or an SP6 promoter.

11. The nucleic acid molecule of claim 9 , wherein the RNA molecule is a self-replicating RNA molecule.

12. The nucleic acid molecule of claim 9 , wherein the RNA molecule further comprises a 5′ cap having a Cap 1 structure, a Cap 1 ( m6 A) structure, a Cap 2 structure, or a Cap 0 structure.

13. A composition comprising the nucleic acid molecule of claim 1 and a lipid formulation selected from a lipoplex, a liposome, a lipid nanoparticle, a polymer-based carrier, an exosome, a lamellar body, a micelle, and an emulsion.

14. The composition of claim 13 , wherein the lipid formulation is a lipid nanoparticle.

15. The composition of claim 13 , wherein the lipid formulation comprises an ionizable cationic lipid.

16. The composition of claim 15 , wherein the ionizable cationic lipid has a structure of Formula I:

or a pharmaceutically acceptable salt or solvate thereof, wherein R 5 and R 6 are each independently selected from the group consisting of a linear or branched C 1 -C 31 alkyl, C 2 -C 31 alkenyl or C 2 -C 31 alkynyl and cholesteryl; L 5 and L 6 are each independently selected from the group consisting of a linear C 1 -C 20 alkyl and C 2 -C 20 alkenyl; X5 is —C(O)O—, whereby —C(O)O—R6 is formed or —OC(O)— whereby —OC(O)—R 6 is formed; X 6 is —C(O)O— whereby —C(O)O—R 5 is formed or —OC(O)— whereby —OC(O)—R 5 is formed; X 7 is S or O; L 7 is absent or lower alkyl; R 4 is a linear or branched C 1 -C 6 alkyl; and R 7 and R 8 are each independently selected from the group consisting of a hydrogen and a linear or branched C 1 -C 6 alkyl.

17. The composition of claim 15 , wherein the ionizable cationic lipid has a structure of:

or a pharmaceutically acceptable salt thereof.

18. The composition of claim 13 , wherein the lipid formulation encapsulates the nucleic acid molecule or is complexed to the nucleic acid molecule.

19. The composition of claim 13 , wherein the lipid formulation comprises

(a) a helper lipid;

(b) a phospholipid;

(c) a polyethylene glycol (PEG)-lipid conjugate; or

(d) any combination thereof.

20. The composition of claim 19 , wherein the phospholipid is selected from dioleoylphosphatidyl ethanolamine (DOPE), dimyristoylphosphatidyl choline (DMPC), distearoylphosphatidyl choline (DSPC), dimyristoylphosphatidyl glycerol (DMPG), dipalmitoyl phosphatidylcholine (DPPC), and phosphatidylcholine (PC).

21. The composition of claim 13 , wherein the lipid portion of the lipid formulation comprises about 40 mol % to about 60 mol % of the ionizable cationic lipid, about 4 mol % to about 16 mol % DSPC, about 30 mol % to about 47 mol % cholesterol, and about 0.5 mol % to about 3 mol % PEG2000-DMG.

22. The composition of claim 13 , wherein the composition has a total lipid: nucleic acid molecule weight ratio of about 50:1 to about 10:1.

23. The composition of claim 13 , wherein the composition comprises

(a) a HEPES or TRIS buffer at a pH of about 7.0 to about 8.5;

(b) a HEPES or TRIS buffer at a concentration of about 7 mg/mL to about 15 mg/ml;

(c) about 2.0 mg/mL to about 4.0 mg/mL of NaCl;

(d) one or more cryoprotectants;

(e) one or more cryoprotectants selected from sucrose, glycerol, or a combination of sucrose and glycerol; or

(f) any combination thereof.

24. The composition of claim 13 , wherein the composition is a lyophilized composition.

25. The composition of claim 24 , wherein the lyophilized composition comprises one or more lyoprotectants.

26. The composition of claim 24 , wherein the lyophilized composition comprises a poloxamer, potassium sorbate, sucrose, or any combination thereof.

27. The composition of claim 24 , wherein the lyophilized composition comprises

(a) about 0.01 to about 1.0% w/w of the nucleic acid molecule;

(b) about 1.0 to about 5.0% w/w lipids;

(c) about 0.5 to about 2.5% w/w of TRIS buffer;

(d) about 0.75 to about 2.75% w/w of NaCl;

(e) about 85 to about 95% w/w of a sugar;

(f) about 0.01 to about 1.0% w/w of a poloxamer;

(g) about 1.0 to about 5.0% w/w of potassium sorbate; or

(h) any combination thereof.

28. A method of administering the composition of claim 13 to a subject in need thereof, wherein the composition is lyophilized and is reconstituted prior to administration.

29. A method of preventing or ameliorating COVID-19, comprising administering the composition of claim 13 to a subject in need thereof.

30. A method of administering a booster dose to a vaccinated subject, comprising administering the composition of claim 13 to a subject who was previously vaccinated against coronavirus.

31. The method of claim 29 , wherein the composition is administered at a dosage of about 0.01 ug to about 1,000 ug of nucleic acid.

32. A method of inducing an immune response against a coronavirus in a subject comprising:

administering to the subject an effective amount of a nucleic acid molecule of claim 1 .

33. A method of inducing an immune response against a coronavirus in a subject comprising:

administering to the subject an effective amount of a composition of claim 13 .

Assignments (3)

Continuity (4)

References Cited (189)

Cited By (2)

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